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Fermentation process and its contribution in pharmacy
Department of Pharmaceutical Sciences Page 1
Assignment on
FERMENTATION PROCESS
And it’s contribution
In Pharmacy
Submitted By: Himangshu Sharma
Roll No: 05
B.Pharm 5th
Semester
Dept. of Pharmaceutical Sciences,
Dibrugarh University
Fermentation process and its contribution in pharmacy
Department of Pharmaceutical Sciences Page 2
Contents: Page No.
1. Introduction 02
1.1 History of fermentation 03
1.2 Definition 03
1.3 Benefit of fermentation 04
1.4 Effect of food 06
2. Type of fermentation 08
3. Factors affecting fermentation 12
4. Chemical &Pharmaceutical made by fermentation 12
4.1 Production of alcoholic beverage 12
4.2 Ethanol 13
4.3 Industrial enzymes 15
4.4 Vitamins 15
4.5 Fermentation using Antibiotic production 16
4.6 Pharmaceutical produced by direct fermentation 17
4.7 Organic acid fermentation 18
5. Biopolymer 19
6. Reference 24
Fermentation process and its contribution in pharmacy
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1. Introduction:
Fermentation was traditionally a process which enabled to preserve food &as such
has been used for long time. However now a days, the main purpose of food
fermentation isn’t to preserve.
Historically, fermentation products were mainly food products, but in recent years an
increased interest has been changes in the production of bulk chemicals,
pharmaceuticals, biofuels, food additives &agriculture.
Fermentation products include:
Food products-
 From milk-Yogurt, kefir, fresh, cheeses
 From fruits-Wine &Vinegar
 From vegetable-pickles, soy sauce, sauerkraut
Industrial chemical-
Solvents-Acetone, butanol, ethanol
Enzymes
Amino acids
Vitamins
Other pharmaceuticals-Antibiotics etc
The main fermentation products include organic acids, ethyl
alcohol&CO2.Commercially the most important are Lactic acid &ethanolic
fermentation. Lactic acid fermentation is used in fermentation of milk,
vegetables, cereals, meats &fish.
In alcoholic fermentation is one of the most important& oldest processes. It
is used in the production of alcoholic beverages, chemical &automotive
industry, solvents, pharmaceutical industry.[1]
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1.1 History of Fermentation:
Fermentation is a natural process. In the 1850s and 1860s Louis Pasteur became the
first scientist to study fermentation when he demonstrated fermentation was caused
by living cells.
The earliest evidence of an alcoholic beverage, made from fruit, rice, and honey,
dates from 7000–6600 BCE, in the Neolithic Chinese village of and winemaking
dates from 6000 BCE, in Georgia, in the Caucasus area. Seven-thousand-year-old
jars containing the remains of wine have been excavated in the Zagros Mountains in
Iran (Dirar, 1993). There is strong evidence that people were fermenting beverages
in Babylon circa 3000 BC (Sahrhage, 2008) pre-Hispanic Mexico circa 2000 BC,
and Sudan circa 1500 BC. Links between fermented foods and health can be traced
as far back as ancient Rome and China, and remain an area of great interest for
researchers in modern times. Wang and Hesseltine (1979) opined that probably the
first fermentation was discovered accidentally when salt was incorporated with the
food material, and the salt selected certain harmless microorganisms to make the
fermented product a nutritious and acceptable food. Lactic fermentation is an ancient
method used by many people throughout the world for preserving vegetables [2]
1.2 Definition:
The term fermentation comes from the Latin word fermantum (to ferment). The
historical definition describes fermentation as the process in which chemical changes
in an organic substrate occur as the result of action of microbial enzymes.
Fermentation can be described as respiration without air.
Historically, the science of fermentation is called zymology and the first zymologist
was Louis Pastuer, who as the first made yeast responsible for fermentation.
Alchemy called fermentation putrefaction – natural rotting or decomposing of
substances. Nowadays, it is a metabolic process in which carbohydrates and related
compounds are partially oxidized with the release of energy in the absence of any
external electron acceptors – organic compounds produced by breakdown of
carbohydrates. During fermentation incomplete oxidation of organic compounds
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occurs and for this reason less energy is obtained when compared with aerobic
oxidation of the compound.
Paradoxically, the term industrial fermentation usually refers to either aerobic or
anaerobic processes, whereas fermentation in biochemical context describes a
strictly anaerobic process, which occurs if pyruvic acid does not enter the Kreb’s
cycle and if electrons from glucose metabolism do not enter electron transport
system. In this process, reduced organic compounds are formed, usually acid by-
products. Industrial fermentation, a term used in chemical engineering, describes the
process that operates that utilize a chemical change induced by a living organism or
enzyme, in particular bacteria, yeast, molds or fungi produce a specific product.
1.3Benefits of fermentation:
Benefits of fermentation include conversion of sugars &others carbohydrates: juice
in to wine, grain to bear or CO2 to leaven bread, sugars in vegetables to preservative
organic acids. Fermentation
-extends shelf life of foods,
-Adds aroma &flavors
-in some cases increases the content of vitamins& improve digestibility.
It can also reduce or increase the toxicity.[1]
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Table: 1Benefits of fermentation:[1]
Benefits Description
General advantages  Development of unique flavors textures of
food.
 Low consumption of energy.
 Low capital& operating costs.
 Relatively simple technology.
.
Pathogenic bacteria& spoilage
organisms are inhibited
The most food is fermented by lactic acid
fermentation, during which pH is lowered to 4.Also
bacteriocins, hydrogen peroxide, ethanol are produced.
They inhabit the growth of unwanted microorganisms
&prevent spoilage of food.
Detoxification & softening Lactic acid fermentation also may reduce the content
of natural toxins in plant food.e.g.-cyanogenic
glycosides’ in cassava &also soften plant tissue.
Beneficial health effects Fermentation improves food safety quality through the
presence of probiotics that protect from E.coli &other
pathogens &have hypocholesterolemic &anti
carcinogenic effects, which is particular significance
in lactose intolerance& gastrointestinal disorders.
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1.4 Effect of food:
Fermentation of foods is the controlled action of microorganisms to alter the texture
of food &to preserve (by the production of acids & alcohols) & to produce
characteristic flavors &aromas.
Changes produced by fermentation in food are discussed in below table: 2 [1]
Change Description
Texture Food is softened as result of complex changes in proteins &
carbohydrates.
Nutritional value Microorganisms improve digestibility by hydrolysis of polymeric
compounds, mainly polysaccharides & proteins; secrete e.g.-
vitamins.
Enrichment with Protein, essential amino acids, essential fatty acids.
Flavor Sugars are fermented to acids, which reduce sweetness & increase
acidity, in some cases bitterness is reduced by enzymatic activity.
Aroma The production of volatile compounds: amines, fatty acids,
aldehydes, esters &ketones.
Color Proteolytic activity, degradation chlorophyll &enzymatic
browning may produce brown pigments.
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Figure 1 Different parts of a fermenter
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2. Type of Fermentation
The most important types of fermentation are as follows:
1. Solid State Fermentation
2. Submerged Fermentation
3. Anaerobic Fermentation
4. Aerobic Fermentation
1. Solid State Fermentation:
In such fermentations, microbial growth and product formation occur at the surface
of solid substrates. Examples of such fermentations are mushroom cultivation, mold-
ripened cheeses, starter cultures, etc. More recently, this approach has been used for
the production of extracellular enzymes, certain valuable chemicals, fungal toxins,
and fungal spores (used for biotransformation).
The substrate provides a rich and complex source of nutrients, which may or may
not need to be supplemented. Such substrates selectively support mycelial
organisms, which can grow at high nutrient concentrations and produce a variety of
extracellular enzymes, e.g., a large number of filamentous fungi, and a few bacteria
(actinomycetes and one strain of Bacillus).
According to the physical state, solid state fermentations are divided into the
following two groups:
(i) Low moisture solids fermented without or with occasional/continuous
agitation.
(ii) Suspended solids fermented in packed columns through which liquid
is circulated.
Solid state fermentations offer certain unique advantages but suffer from some
important disadvantages. Products and waste products and cells are continuously
removed for processing
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2. Submerged Fermentation:
(i) Batch Culture:
Batch culture is a closed culture system, which contains limited amount of nutrient
medium. After inoculation, the culture enters lag phase, during which there is
increase in the size of the cells and not in their number. The culture then enters lag
phase or exponential growth phase during which cells divide at a maximal rate and
their generation time reaches minimum.
The increasing population of bacterial cells, after sometime, enters into a stationary-
phase due to depletion of the nutrients and the accumulation of inhibitory end
products in the medium. Eventually, the stationary, phase of bacterial population
culminates into death-phase when the viable bacterial cells begin to die.
(ii) Fed-Batch Culture:
When a butch culture is subsequently led with fresh nutrient medium without
removing the growing microbial culture, it is called fed-batch culture. Fed-batch
culture allows one to supplement the medium with such nutrients that are depleted or
that may be needed for the terminal stages of the culture, e.g., production of
secondary metabolites.
Therefore, the volume of a fed- batch culture increases with time. Fed-batch cultures
achieve higher cell densities than batch cultures. It is used when high substrate
concentration causes growth inhibition. It allows the substrate to be used at lower
non-toxic levels, followed by subsequent feeding. It allows the maximum production
of cellular melabolities by the culture.
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(iii)Continuous Culture:
Where fresh media is continuously added and bioreactor fluid is continuously
removed. As a result, cells continuously receive fresh medium or nutrients and end
products are continuously removed. A continuous culture, the growth of bacterial
population can be maintained in a steady state over a long period of time.
The reactor can thus be operated for long periods of time without having to be shut
down.
3. Anaerobic Fermentation:
In anaerobic fermentation, a provision for aeration is usually not needed. But in
some cases, aeration may be needed initially for inoculum build-up. In most cases, a
mixing device is also unnecessary, but in some cases initial mixing of the inoculum
is necessary. Once the fermentation begins, the gas produced in the process
generates sufficient mixing.
The air present in the headspace of the fermenter should be replaced by CO2, H2, N2
or a suitable mixture of these; this is particularly important for obligate anaerobes
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like Clostridium. The fermentation usually liberates CO2 and H2, which are collected
and used, e.g., CO2 for making dry ice and methanol, and for bubbling into freshly
inoculated fermenters.
Recovery of products from anaerobic fermenters does not require anaerobic
conditions. But many enzymes of such organisms are highly 02-sensitive. Therefore,
when recovery of such enzymes is the objective, cells must be harvested under
strictly anaerobic conditions.
4. Aerobic Fermentation:
The main feature of aerobic fermentation is the provision for adequate aeration; in
some cases, the amount of air needed per hour is about 60-times the medium
volume. Therefore, bioreactors used for aerobic fermentation have a provision for
adequate supply of sterile air, which is generally sparged into the medium. In
addition, these fermenters may have a mechanism for stirring and mixing of the
medium and cells.
Aerobic fermenters may be either of the
(i) stirred-tank type in which mechanical motor-driven stirrers are provided
or
(ii) Of air-lift type in which no mechanical stirrers are used and the
agitation is achieved by the air bubbles generated by the air supply.
Generally, these bioreactors are of closed or batch types, but continuous
flow reactors are also used; such reactors provide a continuous source of
cells and are also suitable for product generation when the product is
released into the medium.
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3. Factor affecting the fermentation processes:
A fermentation is influenced by numerous factors, including temperature, pH, nature
and composition of the medium, dissolved oxygen, dissolved carbondioxide,
operational system (eg.: batch, fed-batch, continuous) feeding with precursors,
mixing (cycling through varying environments), and shear rates in the fermenter.
Variations in these factors may affect : the rate of fermentation; the product
spectrum and yield; the organoleptic properties of the product (appearance, taste,
smell and texture); the generation of toxins; nutritional quality; and other physio-
chemical properties.
The formulation of the fermentation medium affects the yield, rate and product
profile. The medium must provide the necessary amounts of carbon, nitrogen, trace
elements and micronutrients (eg.: vitamins). Specific types of carbon and nitrogen
soueces may be required, and the carbon : nitrogen ratio may have to be controlled.
An understanding of fermentation biochemistry is essential for developing a medium
with an appropriate formulation. Concentrations of certain nutrients may have to be
varied in a specific way during a fermentation to achieve the desired result. Some
trace elements may have to be avoided – for example, minute amounts of iron reduce
yields in citric acid production by Aspergillus niger. Additional factors, such as
cost, availability, and batch-to-batch variability also affect the choice of
medium.[11]
4. Chemical &Pharmaceutical made by fermentation:
4.1 Production of alcoholic beverage:
The fermentation of juices of grapes, cherries and berries to produce wine is an old
and well established procedure. In this instance, the sugar from these juices may be
converted to alcohol via the Embden-Meyerhof pathway. Flavors and aromas occur
in the wine as a result of the activities of chemical reactions other than sugars that
proceed at the same time.
The organisms used in the production of wine are also important. The type of
organisms used will influence the quality of the wine, and a variety of different
species are used, some to produce especially sweet or dry wine. Traditionally, the
yeasts used to ferment the juices were those occurring naturally on the surface of the
fruit. Recently, the trend has been utilizing laboratory cultures of strains of
Saccharomyces cerevisiae var. ellipsoideus. The strains are chosen for their ability to
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properly ferment the variety of grapes used and for their ability to impart flavors by
formation of by-products which a characteristics for a particular wine.
Temperature is especially important as the yield of alcohol is higher at the lower
temperatures and it also encourages the formation of pleasant flavors. At higher
temperatures the alcohol resistance of the yeast is decreased. A favorable
temperature for fermentation is in the vicinity of 100
C.
Beer:
Beer is a alcoholic beverage prepared from fermented grains, usually barley. Several
different starting materials may be used in the production of beer, but they all
achieve the same end – the production of a carbonated alcoholic drink. The top
fermenting yeast, Saccharomyces cerevisiae, is the most widely used among all the
yeast. Strains should be chosen that are low-temperature tolerant varieties. In this
manner, the low temperature favors the growth of the yeast and not bacteria, which
may enter as contaminants on this starting material. Not all beer is made form
Saccharomyces cerevisiae. For example, Saccharomyces carlbergensis and
Saccharomyces monacensis are used. These yeasts grow at the bottom and are
known as bottom fomenters.
Whisky:
They are made from fermented grains (Corn, wheat, barley malt or rye malt), which
are mixed in varying proportion according to the type of whisky being produced and
fermented by yeast. The procedure are similar to those involved in beer production,
the major exception is that the fermented grain broth is distilled in order to
concentrate the alcohol.
4.2 Ethanol:
Ethanol is a primary alcohol with many industrial uses. It can be produced from
sugar containing feedstock by fermentation. Alcoholic fermentation is one of the
oldest and most important examples of industrial fermentation. Traditionally, this
process has been used to produce alcoholic beverages, but today it also plays an
outstanding role in the chemical and automotive industry. Ethanol is also an
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important solvent and starting material for cosmetics and pharmaceuticals and is also
widely used as a disinfectant in medicine.
Ethanol is produced from carbohydrate materials by yeasts in an extra-cellular
process.
I. Feedstock preparation: Sugarcane or sorghum must be crushed to extract their
simple sugars. Starches are converted to sugars in two stages, liquefaction and
saccharification, by adding water, enzymes, and heat (enzymatic hydrolysis).
II. Fermentation: The mash is transferred to the fermentation tank and cooled to the
optimum temperature (around 30 °C).Care has to be taken to assure that no infection
(other organisms that compete with the yeast for the glucose) occurs. Then the
appropriate proportion of yeast is added. The yeast will begin producing alcohol up
to a concentration of 8-12 percent and then become inactive as the alcohol content
becomes too high.
Separation:
The mash is now ready for distillation. A simple one step “stripper” distillation
separates the liquid from the solids. The residue of this distillation is a slurry
consisting of the microbial biomass and water, called stillage.It is removed to
prevent clogging problems during the next step, fractionated distillation. It is often
used to produce secondary products, such as animal feed additives or seasonings or it
is converted to methane and burned as an energy source.
III. Distillation: Distillation separates the ethanol from the water in a rectifying
column. The product is 96 % ethanol. It cannot be further enriched by distillation
because of azeotrope formation, but must be dehydrated by other means
IV. Dehydration: Anhydrous ethanol is required for blending gasoline. It can be
obtained by additional dehydration, e.g. with molecular sieves or carrier assisted
distillation [7].
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4.3 Industrial enzymes:
Of primary interest among the intracellular components are
microbial enzymes: catalase, amylase, protease, pectinase, glucose
isomerase, cellulase, hemicellulase, lipase, lactase, streptokinase and many others.
Recombinant proteins, such as insulin, hepatitis B vaccine, interferon, granulocyte
colony-stimulating factor, streptokinase and others are also made this way.
4.4 Vitamins:
Vitamins are produced by fermentation of sugar containing starting materials and
special additives by bacteria or yeast. They are produced inside the cell and not
released into the fermentation broth. The process parameters are similar to same as
alcohol; the difference being the additives, which are essential components of the
vitamins.
 Vitamin A1 (retinal) is produced from β-carotene, which can be obtained by
fermentation of corn, soybean meal, kerosene, thiamin and α-ionone. The
dry-mass after fermentation contains 120 –150 g product/kg.
 Vitamin B2 (riboflavin) is produced by yeast from glucose, urea and mineral
salts in an aerobic fermentation.
 Vitamin B12 (cyanocobalamine) is produced by bacteria from glucose, corn
and cobalt salts in anaerobic (3 days) and then an aerobic fermentation (also
3 days).
 The starting point for synthesis of Vitamin C is the selective of oxidation of
the sugar compound D-sorbit to L-sorbose using Acetobacter suboxidans
bacteria. L-sorbose is then converted to L-ascorbic acid, better known as
Vitamin C.
 Vitamin D2 is formed by photochemical cleavage of ergosterin,which is a
side product of many fermentation processes. Microorganisms usually
contain up to 3 % of ergosterin[7].
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4.5 Fermentation using Antibiotic production:
Although most research is devoted to the biological and pharmacological problems,
the key step in the actual production of biotech pharmaceuticals is fermentation.
This is demonstrated by the examples, penicillin, insulin, interferon, and
erythropoietin (EPO) – to name just a few.
Penicillin changed the world! It was the first highly efficient antibiotic
pharmaceutical that allowed an effective treatment of bacterial infections.
Penicillin was discovered in 1928 by Alexander Fleming by chance. He observed
that the growth of a bacteria culture was inhibited by a fungus Penicillum notatum.
Penicillin did not only change the medical world, but also the fermentation
technology. The naturally growing (wild type) Penicillum notatum produced
penicillin with a yield of 10 mg/l.
To enhance the penicillin production further, the old method of growing the
Penicillum mold on the surface of the medium in liter-sized flasks was replaced by
fermentation in large aerated tanks.This allowed the mold to grow throughout the
entire tank and not just on the surface of the medium.
Today, penicillin and other antibiotics are produced in large scale fermenters holding
several hundred cubic meters of medium and the yield has increased 5000 fold to 50
g/l.
Equation shows a simplified scheme of the biosynthesis of penicillin. It starts with
the amino acids L-α-aminoadipic acid and L-cysteine from penicillin N in a complex
reaction sequence. When phenyl acetic acid is added to the fermentation medium,
the side chain of the molecule is modified and the
resulting product is called penicillin G[10].
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4.6 Pharmaceutical produced by biotransformation:
Bio transformations are chemical reactions that are induced by enzymes in the cells.
Sometimes it is possible to isolate the enzymes and to carry out the chemical
reaction in a separate reactor in the absence of living cells.
Starting materials are single chemical compounds or mixtures of related compounds,
which are converted to the product with high selectivity. Many biotransformations
are difficult to achieve by conventional synthesis. A classical example is the
synthesis of chiral molecules.
A compound is chiral, when can occur in two forms that are mirror images of each
other. Classical synthesis produces both enantiomers in a 1 to 1 ratio. They cannot be
separated by normal physical means. Nature is, however, more selective. Here only
single enantiomers are formed. This can be utilized to separate D, L enantiomers of
amino acids.
The enzyme L-amylase produces selectively the Lamino acid from a mixture of the
DL-acylamino acids.
A compound is chiral, when can occur in two forms that are mirror images of each
other. Classical synthesis produces both enantiomers in a 1 to 1 ratio. They cannot be
separated by normal physical means. Nature is, however, more selective. Here only
single enantiomers are formed. This can be utilized to separate D, L enantiomers of
amino acids. The enzyme L-amylase produces selectively the L-amino acid from a
mixture of the DL-acylamino acids [7]
The same compound is converted to the amino acid L(+)-aspartic acid by
Escherichia bacteria that contain the enzyme aspartase. If Pseudomonas bacteria are
added, another amino acid L-alanine is formed [7].
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4.7 Organic acid fermentation
Following are the organic acid prepared by fermentation process:
1. Citric acid
2. Gluconic acid
3. D-Lactic acid
4. Gallic acid
5. Fumeric acid
6. Itaconic acid
7. Glycerol
1. Citric Acid:
Citric acid is the product of fermentation of numerous organisms. However, certain
strains of the fungus Aspergillus niger produced commercially high yield of citric
acid from a variety of 2-, 3-, 4-, 5-, 7- or 12-carbon compounds.
Uses: Commercially as a flavoring ingredient in beverage and foods,
especially in dye mixtures such as gelatins and soft drink powders as tablets and as
the principle acid in the preparation of soft drinks, desserts, jams, jellies, candies,
wines and frozen fruits. The acid is rapidly and most commonly metabolized in the
human body and has wide pharmaceutical uses. Especially its incorporation is
effervescent product and as citrates in blood transfusion. Citric acid is used in
astringent lotion to adjust the pH, in hair rinses and hair setting preparation, and
electro plating, in leather tanning, and in inactivating clogged with iron [10].
2. Gluconic Acid
It is formed from sugars by the action of large number of species of molds, chiefly
species of Aspergillus and penicillium [10].
3. D-Lactic Acid
D-Lactic is produced from the fungus Rhizopus oryzae, it is a rapid process using a
rotary fermenter with force aeration. The time of the fermentation was reduced to
30-35 hours and yield of 70-75% were claimed [10]
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4. Gallic Acid
It was obtained by fermenting a clear extract of tanning by means of an organism
which he named Aspergillus gallomyces [10].
5. Fumeric Acid
It is produced by fermentation of sugar with the help of Rhizopus arrhizus. The aicd
is required chiefly for the manufacture of plastics and varnishes [10].
6. Itaconic Acid
It was first obtained as a mould metabolic product of Aspergillus itaconicus [10].
7. Glycerol
Glycerol is formed in a small amount during the normal alcoholic fermentation of
sugar by yeast [10].
5. Pharmaceutical Products by Fermentation Biopolymers:
Biopolymers are polymers produced by living organisms; in other words, they are
polymeric biomolecules, biopolymers contain monomeric units that are covalently
bonded to form larger structures.
There are three main classes of biopolymers-
 Polynucleotides (RNA and DNA): which are long polymers composed of 13
or more nucleotide monomers.
 Polypeptides: which are short polymers of amino acids.
 Polysaccharides: which are often linear bonded polymeric carbohydrate
structures.[3][4][5][6]
Cellulose is the most common organic compound and biopolymer on Earth.
Many membranes, proteins, and nucleotides that are present in living organisms are
polymers. Industrial biopolymers are still niche products, but they are gaining
rapidly in importance, since they have advantages in special applications.
 Here are a few examples: Water-soluble carbohydrate
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Other important aspects are that polysaccharides come from natural, renewable
sources, that they are bio-compatible and biodegradable.
 For example, Xanthan gum is a water soluble heteropolysaccharide with a
very high molecular weight (> 1 million) produced by the bacterium
Xanthomonas campestris.
It is used in food processing as a stabilizer for sauces and dressings.
 Biopolymers are also used in adhesives, water color, printing inks,cosmetics,
and in the pharmaceutical industry.
Polylactides are made from lactic acid and are use for orthopedic repair materials.
 They can be molded or converted into films, fibers, and non-woven fabrics.
The biopolymer is produced by low-cost fermentation or from waste streams
substrates.Polyhydroxyalkanoic acids (PHAs) have been extensively
researched since the 1970s because of the potential applications.The most
successful PHA products are the polyhydroxybutyrates.[7]
 Role of Biopolymers in Green Nanotechnology:
The biopolymer matrix offers additional advantages like water solubility and
biocompatibility necessary for use in biological applications. Hence, the
starch capped water soluble nanoparticles exhibited excellent antibacterial
activity against both gram positive and gram negative bacteria at a very low
concentration.
Polymers like polyvinyl pyrollidone (PVP) and polyacrylamide have been
successfully used as the stabilizing agents for synthesis of various metal
nanoparticles.
 Biopolymers in Drug Delivery:
 Cellulose and its derivatives:
Sarch, carboxymethyl cellulose (CMC), methyl cellulose (MC),
hydroxyethyl cellulose (HEC), hydroxypropyl methyl cellulose (HPMC),
hydroxypropyl cellulose (HFC), ethyl hydroxyethyl cellulose (EHEC),
and methyl hydroxyethyl cellulose (MHEC) are used as an excipient in
many different types of dosage forms [9].
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 Pectin:
Pectin has been investigated as an excipient in many different types of
dosage forms such as filmcoating of colon-specific drug delivery systems
when mixed with ethyl cellulose, Microparticulate delivery systems for
ophthalmic preparations and matrix type transdermal patches.
It has high potential as a hydrophilic polymeric material for controlled
release matrix drug delivery systems, but its aqueous solubility
contributes to premature and fast release of the drug from these matrices
[9].
 Chitin and its derivatives:
Chitin, chitosan, and their derivatives have found a number of
pharmaceutical or biomedical applications.Although chitosan has been
mostly used as a diluent in tablet manufacturing.
It has been also proposed as a binder, lubricant, or potential disintegrating
agent.
The mucoadhesive properties of chitosan make it an attractive material for
the local delivery of drugs in the oral cavity [9].
 Alginates:
It has been used as stabilizers in emulsions, suspending agents, tablet
binders and tablet disintegrators [9].
 Gums and Mucilage:
sVarious gums and mucilages were used in various forms as sustained
release excipient, binder, disintegrant etc.
(E.g-Xanthane gum, Guar gum etc.)[9].
.
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6. References:-
1.‘Fermentation product’ Chemical Engineering &Chemical process Technology-
Vol.-V, by-K.Chojnacka, Institute of Inorganic Technology &Mineral Fertilizers,
Wroclaw Technology, Poland.
2. TRADITION, TREND AND PROSPECT OF FERMENTED FOOD
PRODUCTS: A BRIEF OVERVIEW.’ Rina Rani Ray1*and Sonali Roy2
1Postgraduate Department of Zoology, Bethune College, 181 Bidhan Sarani,
Kolkata.
2Department of History, Jogesh Chandra Chaudhuri College, 30 Prince Anwar Shah
Road, Kolkata.
3.Mohanty, A.K., et al., Natural Fibers, Biopolymers, and Biocomposites (CRC
Press, 2005)
4.Chandra, R., and Rustgi, R., "Biodegradable Polymers", Progress in Polymer
Science, Vol. 23, p. 1273 (1998)
5.Meyers, M.A., et al., "Biological Materials: Structure & Mechanical Properties",
Progress in Materials Science, Vol. 53, p. 1 (2008)
6. Kumar, A., et al., "Smart Polymers: Physical Forms & Bioengineering
Applications", Progress in Polymer Science, Vol. 32, p.1205 (2007)
7. Chapter-9 in industrial microbiology
8. Role of Biopolymers in Green Nanotechnology Sonal I. Thakore Department of
Chemistry, Faculty of Science, The Maharaja Sayajirao University of
Baroda,Vadodara, Gujarat India;120-132.
9. Pharmacologyonline 1: 666-674 (2011) newsletter Akhilesh V Singh 666;
Biopolymers in Drug Delivery: A Review Akhilesh V Singh Department of
Pharmaceutical Sciences, Dibrugarh University, Dibrugarh, Assam, India; 666-673.
10.”.Enzyme- base Industrial Fermentation Technology” in Biotechnology
Fundamentals & Applications by S.S.Puruhit ,Published by Agrobios(India) agro
House,Behind Narsari Cinema,Jodpur-342002,4th edition,page no.-841-852.
11. Christi, Y., in Encyclopedia of Food Microbiology, Robinson, R., Batt, C., and
Patel, P., editors, Academic press, London, 1999. Pp. 663-674. Fermentation
(industrial): Basic considerations.

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fermentation process &its contribution in pharmacy.

  • 1. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 1 Assignment on FERMENTATION PROCESS And it’s contribution In Pharmacy Submitted By: Himangshu Sharma Roll No: 05 B.Pharm 5th Semester Dept. of Pharmaceutical Sciences, Dibrugarh University
  • 2. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 2 Contents: Page No. 1. Introduction 02 1.1 History of fermentation 03 1.2 Definition 03 1.3 Benefit of fermentation 04 1.4 Effect of food 06 2. Type of fermentation 08 3. Factors affecting fermentation 12 4. Chemical &Pharmaceutical made by fermentation 12 4.1 Production of alcoholic beverage 12 4.2 Ethanol 13 4.3 Industrial enzymes 15 4.4 Vitamins 15 4.5 Fermentation using Antibiotic production 16 4.6 Pharmaceutical produced by direct fermentation 17 4.7 Organic acid fermentation 18 5. Biopolymer 19 6. Reference 24
  • 3. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 3 1. Introduction: Fermentation was traditionally a process which enabled to preserve food &as such has been used for long time. However now a days, the main purpose of food fermentation isn’t to preserve. Historically, fermentation products were mainly food products, but in recent years an increased interest has been changes in the production of bulk chemicals, pharmaceuticals, biofuels, food additives &agriculture. Fermentation products include: Food products-  From milk-Yogurt, kefir, fresh, cheeses  From fruits-Wine &Vinegar  From vegetable-pickles, soy sauce, sauerkraut Industrial chemical- Solvents-Acetone, butanol, ethanol Enzymes Amino acids Vitamins Other pharmaceuticals-Antibiotics etc The main fermentation products include organic acids, ethyl alcohol&CO2.Commercially the most important are Lactic acid &ethanolic fermentation. Lactic acid fermentation is used in fermentation of milk, vegetables, cereals, meats &fish. In alcoholic fermentation is one of the most important& oldest processes. It is used in the production of alcoholic beverages, chemical &automotive industry, solvents, pharmaceutical industry.[1]
  • 4. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 4 1.1 History of Fermentation: Fermentation is a natural process. In the 1850s and 1860s Louis Pasteur became the first scientist to study fermentation when he demonstrated fermentation was caused by living cells. The earliest evidence of an alcoholic beverage, made from fruit, rice, and honey, dates from 7000–6600 BCE, in the Neolithic Chinese village of and winemaking dates from 6000 BCE, in Georgia, in the Caucasus area. Seven-thousand-year-old jars containing the remains of wine have been excavated in the Zagros Mountains in Iran (Dirar, 1993). There is strong evidence that people were fermenting beverages in Babylon circa 3000 BC (Sahrhage, 2008) pre-Hispanic Mexico circa 2000 BC, and Sudan circa 1500 BC. Links between fermented foods and health can be traced as far back as ancient Rome and China, and remain an area of great interest for researchers in modern times. Wang and Hesseltine (1979) opined that probably the first fermentation was discovered accidentally when salt was incorporated with the food material, and the salt selected certain harmless microorganisms to make the fermented product a nutritious and acceptable food. Lactic fermentation is an ancient method used by many people throughout the world for preserving vegetables [2] 1.2 Definition: The term fermentation comes from the Latin word fermantum (to ferment). The historical definition describes fermentation as the process in which chemical changes in an organic substrate occur as the result of action of microbial enzymes. Fermentation can be described as respiration without air. Historically, the science of fermentation is called zymology and the first zymologist was Louis Pastuer, who as the first made yeast responsible for fermentation. Alchemy called fermentation putrefaction – natural rotting or decomposing of substances. Nowadays, it is a metabolic process in which carbohydrates and related compounds are partially oxidized with the release of energy in the absence of any external electron acceptors – organic compounds produced by breakdown of carbohydrates. During fermentation incomplete oxidation of organic compounds
  • 5. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 5 occurs and for this reason less energy is obtained when compared with aerobic oxidation of the compound. Paradoxically, the term industrial fermentation usually refers to either aerobic or anaerobic processes, whereas fermentation in biochemical context describes a strictly anaerobic process, which occurs if pyruvic acid does not enter the Kreb’s cycle and if electrons from glucose metabolism do not enter electron transport system. In this process, reduced organic compounds are formed, usually acid by- products. Industrial fermentation, a term used in chemical engineering, describes the process that operates that utilize a chemical change induced by a living organism or enzyme, in particular bacteria, yeast, molds or fungi produce a specific product. 1.3Benefits of fermentation: Benefits of fermentation include conversion of sugars &others carbohydrates: juice in to wine, grain to bear or CO2 to leaven bread, sugars in vegetables to preservative organic acids. Fermentation -extends shelf life of foods, -Adds aroma &flavors -in some cases increases the content of vitamins& improve digestibility. It can also reduce or increase the toxicity.[1]
  • 6. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 6 Table: 1Benefits of fermentation:[1] Benefits Description General advantages  Development of unique flavors textures of food.  Low consumption of energy.  Low capital& operating costs.  Relatively simple technology. . Pathogenic bacteria& spoilage organisms are inhibited The most food is fermented by lactic acid fermentation, during which pH is lowered to 4.Also bacteriocins, hydrogen peroxide, ethanol are produced. They inhabit the growth of unwanted microorganisms &prevent spoilage of food. Detoxification & softening Lactic acid fermentation also may reduce the content of natural toxins in plant food.e.g.-cyanogenic glycosides’ in cassava &also soften plant tissue. Beneficial health effects Fermentation improves food safety quality through the presence of probiotics that protect from E.coli &other pathogens &have hypocholesterolemic &anti carcinogenic effects, which is particular significance in lactose intolerance& gastrointestinal disorders.
  • 7. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 7 1.4 Effect of food: Fermentation of foods is the controlled action of microorganisms to alter the texture of food &to preserve (by the production of acids & alcohols) & to produce characteristic flavors &aromas. Changes produced by fermentation in food are discussed in below table: 2 [1] Change Description Texture Food is softened as result of complex changes in proteins & carbohydrates. Nutritional value Microorganisms improve digestibility by hydrolysis of polymeric compounds, mainly polysaccharides & proteins; secrete e.g.- vitamins. Enrichment with Protein, essential amino acids, essential fatty acids. Flavor Sugars are fermented to acids, which reduce sweetness & increase acidity, in some cases bitterness is reduced by enzymatic activity. Aroma The production of volatile compounds: amines, fatty acids, aldehydes, esters &ketones. Color Proteolytic activity, degradation chlorophyll &enzymatic browning may produce brown pigments.
  • 8. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 8 Figure 1 Different parts of a fermenter
  • 9. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 9 2. Type of Fermentation The most important types of fermentation are as follows: 1. Solid State Fermentation 2. Submerged Fermentation 3. Anaerobic Fermentation 4. Aerobic Fermentation 1. Solid State Fermentation: In such fermentations, microbial growth and product formation occur at the surface of solid substrates. Examples of such fermentations are mushroom cultivation, mold- ripened cheeses, starter cultures, etc. More recently, this approach has been used for the production of extracellular enzymes, certain valuable chemicals, fungal toxins, and fungal spores (used for biotransformation). The substrate provides a rich and complex source of nutrients, which may or may not need to be supplemented. Such substrates selectively support mycelial organisms, which can grow at high nutrient concentrations and produce a variety of extracellular enzymes, e.g., a large number of filamentous fungi, and a few bacteria (actinomycetes and one strain of Bacillus). According to the physical state, solid state fermentations are divided into the following two groups: (i) Low moisture solids fermented without or with occasional/continuous agitation. (ii) Suspended solids fermented in packed columns through which liquid is circulated. Solid state fermentations offer certain unique advantages but suffer from some important disadvantages. Products and waste products and cells are continuously removed for processing
  • 10. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 10 2. Submerged Fermentation: (i) Batch Culture: Batch culture is a closed culture system, which contains limited amount of nutrient medium. After inoculation, the culture enters lag phase, during which there is increase in the size of the cells and not in their number. The culture then enters lag phase or exponential growth phase during which cells divide at a maximal rate and their generation time reaches minimum. The increasing population of bacterial cells, after sometime, enters into a stationary- phase due to depletion of the nutrients and the accumulation of inhibitory end products in the medium. Eventually, the stationary, phase of bacterial population culminates into death-phase when the viable bacterial cells begin to die. (ii) Fed-Batch Culture: When a butch culture is subsequently led with fresh nutrient medium without removing the growing microbial culture, it is called fed-batch culture. Fed-batch culture allows one to supplement the medium with such nutrients that are depleted or that may be needed for the terminal stages of the culture, e.g., production of secondary metabolites. Therefore, the volume of a fed- batch culture increases with time. Fed-batch cultures achieve higher cell densities than batch cultures. It is used when high substrate concentration causes growth inhibition. It allows the substrate to be used at lower non-toxic levels, followed by subsequent feeding. It allows the maximum production of cellular melabolities by the culture.
  • 11. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 11 (iii)Continuous Culture: Where fresh media is continuously added and bioreactor fluid is continuously removed. As a result, cells continuously receive fresh medium or nutrients and end products are continuously removed. A continuous culture, the growth of bacterial population can be maintained in a steady state over a long period of time. The reactor can thus be operated for long periods of time without having to be shut down. 3. Anaerobic Fermentation: In anaerobic fermentation, a provision for aeration is usually not needed. But in some cases, aeration may be needed initially for inoculum build-up. In most cases, a mixing device is also unnecessary, but in some cases initial mixing of the inoculum is necessary. Once the fermentation begins, the gas produced in the process generates sufficient mixing. The air present in the headspace of the fermenter should be replaced by CO2, H2, N2 or a suitable mixture of these; this is particularly important for obligate anaerobes
  • 12. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 12 like Clostridium. The fermentation usually liberates CO2 and H2, which are collected and used, e.g., CO2 for making dry ice and methanol, and for bubbling into freshly inoculated fermenters. Recovery of products from anaerobic fermenters does not require anaerobic conditions. But many enzymes of such organisms are highly 02-sensitive. Therefore, when recovery of such enzymes is the objective, cells must be harvested under strictly anaerobic conditions. 4. Aerobic Fermentation: The main feature of aerobic fermentation is the provision for adequate aeration; in some cases, the amount of air needed per hour is about 60-times the medium volume. Therefore, bioreactors used for aerobic fermentation have a provision for adequate supply of sterile air, which is generally sparged into the medium. In addition, these fermenters may have a mechanism for stirring and mixing of the medium and cells. Aerobic fermenters may be either of the (i) stirred-tank type in which mechanical motor-driven stirrers are provided or (ii) Of air-lift type in which no mechanical stirrers are used and the agitation is achieved by the air bubbles generated by the air supply. Generally, these bioreactors are of closed or batch types, but continuous flow reactors are also used; such reactors provide a continuous source of cells and are also suitable for product generation when the product is released into the medium.
  • 13. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 13 3. Factor affecting the fermentation processes: A fermentation is influenced by numerous factors, including temperature, pH, nature and composition of the medium, dissolved oxygen, dissolved carbondioxide, operational system (eg.: batch, fed-batch, continuous) feeding with precursors, mixing (cycling through varying environments), and shear rates in the fermenter. Variations in these factors may affect : the rate of fermentation; the product spectrum and yield; the organoleptic properties of the product (appearance, taste, smell and texture); the generation of toxins; nutritional quality; and other physio- chemical properties. The formulation of the fermentation medium affects the yield, rate and product profile. The medium must provide the necessary amounts of carbon, nitrogen, trace elements and micronutrients (eg.: vitamins). Specific types of carbon and nitrogen soueces may be required, and the carbon : nitrogen ratio may have to be controlled. An understanding of fermentation biochemistry is essential for developing a medium with an appropriate formulation. Concentrations of certain nutrients may have to be varied in a specific way during a fermentation to achieve the desired result. Some trace elements may have to be avoided – for example, minute amounts of iron reduce yields in citric acid production by Aspergillus niger. Additional factors, such as cost, availability, and batch-to-batch variability also affect the choice of medium.[11] 4. Chemical &Pharmaceutical made by fermentation: 4.1 Production of alcoholic beverage: The fermentation of juices of grapes, cherries and berries to produce wine is an old and well established procedure. In this instance, the sugar from these juices may be converted to alcohol via the Embden-Meyerhof pathway. Flavors and aromas occur in the wine as a result of the activities of chemical reactions other than sugars that proceed at the same time. The organisms used in the production of wine are also important. The type of organisms used will influence the quality of the wine, and a variety of different species are used, some to produce especially sweet or dry wine. Traditionally, the yeasts used to ferment the juices were those occurring naturally on the surface of the fruit. Recently, the trend has been utilizing laboratory cultures of strains of Saccharomyces cerevisiae var. ellipsoideus. The strains are chosen for their ability to
  • 14. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 14 properly ferment the variety of grapes used and for their ability to impart flavors by formation of by-products which a characteristics for a particular wine. Temperature is especially important as the yield of alcohol is higher at the lower temperatures and it also encourages the formation of pleasant flavors. At higher temperatures the alcohol resistance of the yeast is decreased. A favorable temperature for fermentation is in the vicinity of 100 C. Beer: Beer is a alcoholic beverage prepared from fermented grains, usually barley. Several different starting materials may be used in the production of beer, but they all achieve the same end – the production of a carbonated alcoholic drink. The top fermenting yeast, Saccharomyces cerevisiae, is the most widely used among all the yeast. Strains should be chosen that are low-temperature tolerant varieties. In this manner, the low temperature favors the growth of the yeast and not bacteria, which may enter as contaminants on this starting material. Not all beer is made form Saccharomyces cerevisiae. For example, Saccharomyces carlbergensis and Saccharomyces monacensis are used. These yeasts grow at the bottom and are known as bottom fomenters. Whisky: They are made from fermented grains (Corn, wheat, barley malt or rye malt), which are mixed in varying proportion according to the type of whisky being produced and fermented by yeast. The procedure are similar to those involved in beer production, the major exception is that the fermented grain broth is distilled in order to concentrate the alcohol. 4.2 Ethanol: Ethanol is a primary alcohol with many industrial uses. It can be produced from sugar containing feedstock by fermentation. Alcoholic fermentation is one of the oldest and most important examples of industrial fermentation. Traditionally, this process has been used to produce alcoholic beverages, but today it also plays an outstanding role in the chemical and automotive industry. Ethanol is also an
  • 15. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 15 important solvent and starting material for cosmetics and pharmaceuticals and is also widely used as a disinfectant in medicine. Ethanol is produced from carbohydrate materials by yeasts in an extra-cellular process. I. Feedstock preparation: Sugarcane or sorghum must be crushed to extract their simple sugars. Starches are converted to sugars in two stages, liquefaction and saccharification, by adding water, enzymes, and heat (enzymatic hydrolysis). II. Fermentation: The mash is transferred to the fermentation tank and cooled to the optimum temperature (around 30 °C).Care has to be taken to assure that no infection (other organisms that compete with the yeast for the glucose) occurs. Then the appropriate proportion of yeast is added. The yeast will begin producing alcohol up to a concentration of 8-12 percent and then become inactive as the alcohol content becomes too high. Separation: The mash is now ready for distillation. A simple one step “stripper” distillation separates the liquid from the solids. The residue of this distillation is a slurry consisting of the microbial biomass and water, called stillage.It is removed to prevent clogging problems during the next step, fractionated distillation. It is often used to produce secondary products, such as animal feed additives or seasonings or it is converted to methane and burned as an energy source. III. Distillation: Distillation separates the ethanol from the water in a rectifying column. The product is 96 % ethanol. It cannot be further enriched by distillation because of azeotrope formation, but must be dehydrated by other means IV. Dehydration: Anhydrous ethanol is required for blending gasoline. It can be obtained by additional dehydration, e.g. with molecular sieves or carrier assisted distillation [7].
  • 16. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 16 4.3 Industrial enzymes: Of primary interest among the intracellular components are microbial enzymes: catalase, amylase, protease, pectinase, glucose isomerase, cellulase, hemicellulase, lipase, lactase, streptokinase and many others. Recombinant proteins, such as insulin, hepatitis B vaccine, interferon, granulocyte colony-stimulating factor, streptokinase and others are also made this way. 4.4 Vitamins: Vitamins are produced by fermentation of sugar containing starting materials and special additives by bacteria or yeast. They are produced inside the cell and not released into the fermentation broth. The process parameters are similar to same as alcohol; the difference being the additives, which are essential components of the vitamins.  Vitamin A1 (retinal) is produced from β-carotene, which can be obtained by fermentation of corn, soybean meal, kerosene, thiamin and α-ionone. The dry-mass after fermentation contains 120 –150 g product/kg.  Vitamin B2 (riboflavin) is produced by yeast from glucose, urea and mineral salts in an aerobic fermentation.  Vitamin B12 (cyanocobalamine) is produced by bacteria from glucose, corn and cobalt salts in anaerobic (3 days) and then an aerobic fermentation (also 3 days).  The starting point for synthesis of Vitamin C is the selective of oxidation of the sugar compound D-sorbit to L-sorbose using Acetobacter suboxidans bacteria. L-sorbose is then converted to L-ascorbic acid, better known as Vitamin C.  Vitamin D2 is formed by photochemical cleavage of ergosterin,which is a side product of many fermentation processes. Microorganisms usually contain up to 3 % of ergosterin[7].
  • 17. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 17 4.5 Fermentation using Antibiotic production: Although most research is devoted to the biological and pharmacological problems, the key step in the actual production of biotech pharmaceuticals is fermentation. This is demonstrated by the examples, penicillin, insulin, interferon, and erythropoietin (EPO) – to name just a few. Penicillin changed the world! It was the first highly efficient antibiotic pharmaceutical that allowed an effective treatment of bacterial infections. Penicillin was discovered in 1928 by Alexander Fleming by chance. He observed that the growth of a bacteria culture was inhibited by a fungus Penicillum notatum. Penicillin did not only change the medical world, but also the fermentation technology. The naturally growing (wild type) Penicillum notatum produced penicillin with a yield of 10 mg/l. To enhance the penicillin production further, the old method of growing the Penicillum mold on the surface of the medium in liter-sized flasks was replaced by fermentation in large aerated tanks.This allowed the mold to grow throughout the entire tank and not just on the surface of the medium. Today, penicillin and other antibiotics are produced in large scale fermenters holding several hundred cubic meters of medium and the yield has increased 5000 fold to 50 g/l. Equation shows a simplified scheme of the biosynthesis of penicillin. It starts with the amino acids L-α-aminoadipic acid and L-cysteine from penicillin N in a complex reaction sequence. When phenyl acetic acid is added to the fermentation medium, the side chain of the molecule is modified and the resulting product is called penicillin G[10].
  • 18. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 18 4.6 Pharmaceutical produced by biotransformation: Bio transformations are chemical reactions that are induced by enzymes in the cells. Sometimes it is possible to isolate the enzymes and to carry out the chemical reaction in a separate reactor in the absence of living cells. Starting materials are single chemical compounds or mixtures of related compounds, which are converted to the product with high selectivity. Many biotransformations are difficult to achieve by conventional synthesis. A classical example is the synthesis of chiral molecules. A compound is chiral, when can occur in two forms that are mirror images of each other. Classical synthesis produces both enantiomers in a 1 to 1 ratio. They cannot be separated by normal physical means. Nature is, however, more selective. Here only single enantiomers are formed. This can be utilized to separate D, L enantiomers of amino acids. The enzyme L-amylase produces selectively the Lamino acid from a mixture of the DL-acylamino acids. A compound is chiral, when can occur in two forms that are mirror images of each other. Classical synthesis produces both enantiomers in a 1 to 1 ratio. They cannot be separated by normal physical means. Nature is, however, more selective. Here only single enantiomers are formed. This can be utilized to separate D, L enantiomers of amino acids. The enzyme L-amylase produces selectively the L-amino acid from a mixture of the DL-acylamino acids [7] The same compound is converted to the amino acid L(+)-aspartic acid by Escherichia bacteria that contain the enzyme aspartase. If Pseudomonas bacteria are added, another amino acid L-alanine is formed [7].
  • 19. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 19 4.7 Organic acid fermentation Following are the organic acid prepared by fermentation process: 1. Citric acid 2. Gluconic acid 3. D-Lactic acid 4. Gallic acid 5. Fumeric acid 6. Itaconic acid 7. Glycerol 1. Citric Acid: Citric acid is the product of fermentation of numerous organisms. However, certain strains of the fungus Aspergillus niger produced commercially high yield of citric acid from a variety of 2-, 3-, 4-, 5-, 7- or 12-carbon compounds. Uses: Commercially as a flavoring ingredient in beverage and foods, especially in dye mixtures such as gelatins and soft drink powders as tablets and as the principle acid in the preparation of soft drinks, desserts, jams, jellies, candies, wines and frozen fruits. The acid is rapidly and most commonly metabolized in the human body and has wide pharmaceutical uses. Especially its incorporation is effervescent product and as citrates in blood transfusion. Citric acid is used in astringent lotion to adjust the pH, in hair rinses and hair setting preparation, and electro plating, in leather tanning, and in inactivating clogged with iron [10]. 2. Gluconic Acid It is formed from sugars by the action of large number of species of molds, chiefly species of Aspergillus and penicillium [10]. 3. D-Lactic Acid D-Lactic is produced from the fungus Rhizopus oryzae, it is a rapid process using a rotary fermenter with force aeration. The time of the fermentation was reduced to 30-35 hours and yield of 70-75% were claimed [10]
  • 20. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 20 4. Gallic Acid It was obtained by fermenting a clear extract of tanning by means of an organism which he named Aspergillus gallomyces [10]. 5. Fumeric Acid It is produced by fermentation of sugar with the help of Rhizopus arrhizus. The aicd is required chiefly for the manufacture of plastics and varnishes [10]. 6. Itaconic Acid It was first obtained as a mould metabolic product of Aspergillus itaconicus [10]. 7. Glycerol Glycerol is formed in a small amount during the normal alcoholic fermentation of sugar by yeast [10]. 5. Pharmaceutical Products by Fermentation Biopolymers: Biopolymers are polymers produced by living organisms; in other words, they are polymeric biomolecules, biopolymers contain monomeric units that are covalently bonded to form larger structures. There are three main classes of biopolymers-  Polynucleotides (RNA and DNA): which are long polymers composed of 13 or more nucleotide monomers.  Polypeptides: which are short polymers of amino acids.  Polysaccharides: which are often linear bonded polymeric carbohydrate structures.[3][4][5][6] Cellulose is the most common organic compound and biopolymer on Earth. Many membranes, proteins, and nucleotides that are present in living organisms are polymers. Industrial biopolymers are still niche products, but they are gaining rapidly in importance, since they have advantages in special applications.  Here are a few examples: Water-soluble carbohydrate
  • 21. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 21 Other important aspects are that polysaccharides come from natural, renewable sources, that they are bio-compatible and biodegradable.  For example, Xanthan gum is a water soluble heteropolysaccharide with a very high molecular weight (> 1 million) produced by the bacterium Xanthomonas campestris. It is used in food processing as a stabilizer for sauces and dressings.  Biopolymers are also used in adhesives, water color, printing inks,cosmetics, and in the pharmaceutical industry. Polylactides are made from lactic acid and are use for orthopedic repair materials.  They can be molded or converted into films, fibers, and non-woven fabrics. The biopolymer is produced by low-cost fermentation or from waste streams substrates.Polyhydroxyalkanoic acids (PHAs) have been extensively researched since the 1970s because of the potential applications.The most successful PHA products are the polyhydroxybutyrates.[7]  Role of Biopolymers in Green Nanotechnology: The biopolymer matrix offers additional advantages like water solubility and biocompatibility necessary for use in biological applications. Hence, the starch capped water soluble nanoparticles exhibited excellent antibacterial activity against both gram positive and gram negative bacteria at a very low concentration. Polymers like polyvinyl pyrollidone (PVP) and polyacrylamide have been successfully used as the stabilizing agents for synthesis of various metal nanoparticles.  Biopolymers in Drug Delivery:  Cellulose and its derivatives: Sarch, carboxymethyl cellulose (CMC), methyl cellulose (MC), hydroxyethyl cellulose (HEC), hydroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose (HFC), ethyl hydroxyethyl cellulose (EHEC), and methyl hydroxyethyl cellulose (MHEC) are used as an excipient in many different types of dosage forms [9].
  • 22. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 22  Pectin: Pectin has been investigated as an excipient in many different types of dosage forms such as filmcoating of colon-specific drug delivery systems when mixed with ethyl cellulose, Microparticulate delivery systems for ophthalmic preparations and matrix type transdermal patches. It has high potential as a hydrophilic polymeric material for controlled release matrix drug delivery systems, but its aqueous solubility contributes to premature and fast release of the drug from these matrices [9].  Chitin and its derivatives: Chitin, chitosan, and their derivatives have found a number of pharmaceutical or biomedical applications.Although chitosan has been mostly used as a diluent in tablet manufacturing. It has been also proposed as a binder, lubricant, or potential disintegrating agent. The mucoadhesive properties of chitosan make it an attractive material for the local delivery of drugs in the oral cavity [9].  Alginates: It has been used as stabilizers in emulsions, suspending agents, tablet binders and tablet disintegrators [9].  Gums and Mucilage: sVarious gums and mucilages were used in various forms as sustained release excipient, binder, disintegrant etc. (E.g-Xanthane gum, Guar gum etc.)[9]. .
  • 23. Fermentation process and its contribution in pharmacy Department of Pharmaceutical Sciences Page 23 6. References:- 1.‘Fermentation product’ Chemical Engineering &Chemical process Technology- Vol.-V, by-K.Chojnacka, Institute of Inorganic Technology &Mineral Fertilizers, Wroclaw Technology, Poland. 2. TRADITION, TREND AND PROSPECT OF FERMENTED FOOD PRODUCTS: A BRIEF OVERVIEW.’ Rina Rani Ray1*and Sonali Roy2 1Postgraduate Department of Zoology, Bethune College, 181 Bidhan Sarani, Kolkata. 2Department of History, Jogesh Chandra Chaudhuri College, 30 Prince Anwar Shah Road, Kolkata. 3.Mohanty, A.K., et al., Natural Fibers, Biopolymers, and Biocomposites (CRC Press, 2005) 4.Chandra, R., and Rustgi, R., "Biodegradable Polymers", Progress in Polymer Science, Vol. 23, p. 1273 (1998) 5.Meyers, M.A., et al., "Biological Materials: Structure & Mechanical Properties", Progress in Materials Science, Vol. 53, p. 1 (2008) 6. Kumar, A., et al., "Smart Polymers: Physical Forms & Bioengineering Applications", Progress in Polymer Science, Vol. 32, p.1205 (2007) 7. Chapter-9 in industrial microbiology 8. Role of Biopolymers in Green Nanotechnology Sonal I. Thakore Department of Chemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda,Vadodara, Gujarat India;120-132. 9. Pharmacologyonline 1: 666-674 (2011) newsletter Akhilesh V Singh 666; Biopolymers in Drug Delivery: A Review Akhilesh V Singh Department of Pharmaceutical Sciences, Dibrugarh University, Dibrugarh, Assam, India; 666-673. 10.”.Enzyme- base Industrial Fermentation Technology” in Biotechnology Fundamentals & Applications by S.S.Puruhit ,Published by Agrobios(India) agro House,Behind Narsari Cinema,Jodpur-342002,4th edition,page no.-841-852. 11. Christi, Y., in Encyclopedia of Food Microbiology, Robinson, R., Batt, C., and Patel, P., editors, Academic press, London, 1999. Pp. 663-674. Fermentation (industrial): Basic considerations.